Methods and compositions for the treatment of symptoms of prion diseases

ABSTRACT

A therapeutic composition for the treatment of the symptoms of prion diseases and the method for preparing the therapeutic agents is disclosed. The therapeutic composition is a stable pharmaceutical composition comprising one or more digestive and/or pancreatic enzymes. The therapeutic composition may be manufactured by a variety of encapsulation technologies. Delivery of the therapeutic composition may be made orally, through injection, by adherence of a medicated patch or other method. Further, a method of using fecal chymotrypsin level as a biomarker for the presence of a prion disease, or the likelihood of an individual to develop a prion disease is disclosed.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 13/737,225, filed Jan. 9, 2013, now U.S. Pat. No. 9,061,033, whichis a continuation application of U.S. application Ser. No. 12/573,353,filed Oct. 5, 2009, now abandoned, which claims the benefit of U.S.Provisional Application No. 61/102,818, filed Oct. 3, 2008, each ofwhich is incorporated by reference in its entirety herein.

TECHNICAL FIELD

This disclosure relates to a treatment for the symptoms of priondiseases, and more particularly, to the use of pharmaceuticalcompositions comprising one or more digestive enzymes, such as one ormore pancreatic enzymes, in the treatment of the symptoms of priondiseases. The disclosure also relates to a method of makingpharmaceutical compositions comprising one or more digestive enzymes.The disclosure further relates to the use of an individual's fecalchymotrypsin level as a diagnostic marker for determining whether anindividual has a prion disease, as well as to predict whether anindividual will be beneficially treated with the describedpharmaceutical compositions.

BACKGROUND OF THE INVENTION

Dysautonomias can result in symptoms in which one or more areas of thebody are innervated by the autonomic nervous system. While somedysautonomias are well known, other conditions have yet to be determinedas a dysautonomia.

Symptoms of known dysautonomias include: palpitations, chest pain,tachycardia, excessive fatigue, severe fluctuations in blood pressure,excessive sweating, fainting, exercise intolerance, shortness of breath,visual disturbances including blurred vision, tunneling, and doublevision, migraines, dizziness, insomnia, gastrointestinal problemsincluding diarrhea, and constipation, bloody stools, fainting/nearfainting, frequent urination, convulsions, and cognitive impairment.Secondarily others symptoms such as depression, dysthymia, obsessivecompulsive tendencies, and difficulty with ambulation and other symptomsmay also be a part of the dysautonomic picture.

Conditions such as familial dysautonomia (FD), also known also asRiley-Day syndrome, Parkinson's disease, Guillaine-Barre syndrome (GBS),Dopamine-b-Hydroxalase deficiency, baroreflex failure, Guillaine-BarreSyndrome, neuroblastoma and other tumors which affect the neuroendocrinesystem, Aromatic L-Amino Acid Decarboxylase deficiency,Tetrahydrobiopterin deficiency, Familial Paraganglioma syndrome,“Shy-Drager Syndrome,” also referred to as “Multiple System Atrophy” orMSA, Neurally Mediated Syncope, also known as Neurocardiogenic Syncope,fetal fatal insomnia (FFI), diabetic cardiovascular neuropathy,hereditary sensory and autonomic neuropathy type III (HSAN III), Menke'sdisease, monoamine oxidase deficiency states, and other disorders ofdopamine metabolism, dysautonomic syndromes and disorders of thecardiovasular system, Chaga's disease, diabetic autonomic failure, andpure autonomic failure, are well known as conditions associated with orprimarily due to a dysautonomia.

Prion diseases are rare. The general worldwide yearly incidence isapproximately one case per million people. Thus, in the US,approximately 300 de novo cases of sporadic and genetic prion diseaseare observed per year. The genetically transmissible forms of priondisease are about one-tenth as common as the sporadic forms. Thisprevalence is comparable to that observed with the autosomal dominantforms of familial Alzheimer disease and amyotrophic lateral sclerosis(Lou Gehrig's disease).

Prion diseases generally manifest with cognitive difficulties, ataxia,and myoclonus (abrupt jerking movements of muscle groups and/or entirelimbs); however, the order and/or predominance of these features andassociated neurologic and psychiatric findings vary with prion diseasesubtype and/or PRNP mutation. The age at onset ranges from the third toninth decade of life. The course ranges from a few months to severalyears (typically five to seven years, but in rare cases more than tenyears). Death generally results from infection, either by pneumonia(typically from aspiration) or urosepsis. Therapy is aimed atcontrolling symptoms that may cause discomfort. No cure for priondisease currently exists.

The three phenotypes classically associated with genetic prion disease(fCJD, GSS, and FFI), were defined by clinical and neuropathologicfindings long before the molecular basis of this group of disorders wasdiscovered. Although it is now recognized that these three phenotypesare part of a continuum and have overlapping features, it can be helpfulto think of genetic human prion disease at least in part in terms ofthese phenotypes when providing individuals and families withinformation about the expected clinical course.

Familial Creutzfeldt-Jakob Disease (fCJD).

Progressive confusion and memory impairment occur first, followed byataxia and myoclonus. The disease typically manifests between the agesof 30 and 50 years, although a few individuals present before age 30 oras late as the upper 80s. The course from onset to death ranges from afew months to five years. At the endstage of disease, the individual isgenerally bedbound, mute, and immobile, except for myoclonic jerks.

The cognitive impairment observed may initially be mild confusion or itmay be specific for a particular cortical function, such as language orconstructional abilities; however, the resultant picture is one ofglobal dementia. As the disease progresses, neurobehavioral symptoms mayvary considerably. Psychiatric features, including delusions andhallucinations, may also occur.

Ataxia may be either truncal or appendicular, manifesting either as anunsteady gait, clumsiness while carrying out commonly performed tasks(e.g., picking up the salt shaker while dining), or progressivedysarthria. As the ataxia progresses, the individual may fallrepeatedly, necessitating the use of a wheelchair to prevent injury.

Myoclonus generally, but not always, occurs after cognitive impairmentis evident. Myoclonus may begin focally in a single limb but eventuallybecomes generalized. “Startle myoclonus” may be elicited by simple actssuch as clapping the hands or turning on the room lights. Even if warnedof an impending noise, the individual cannot suppress the startleresponse.

Other neurologic signs and symptoms such as focal or generalizedweakness, rigidity, bradykinesia, tremor, chorea, alien hand syndrome,stroke-like symptoms, visual disturbances, and seizures have beenobserved.

Gerstmann-Sträussler-Scheinker Syndrome (GSS).

GSS typically begins in the fourth to sixth decade with the insidiousonset of cerebellar dysfunction, manifest as unsteady gait and milddysarthria. Cognitive dysfunction is generally not apparent early on;however, with progression, bradyphrenia, or slowness of thoughtprocessing, may become evident. Pyramidal involvement with spasticityand/or extrapyramidal involvement with bradykinesia, increased muscletone with or without cogwheeling, and masked facies are also common.Psychiatric or behavioral symptoms are atypical. The disease progressesat a relatively slow but relentless pace over the course of a few toseven or more years. Cerebellar dysfunction results in severedysarthria, gait and appendicular ataxia, ocular dysmetria, and lack ofcoordination in swallowing. A decline in cognitive abilities,particularly of concentration and focus, becomes apparent withprogression into the late stage of disease. In the terminal stage, theindividual is bedridden from the disabling ataxia, unable to eat becauseof severe lack of coordination in swallowing, and unable to communicatebecause of the profound dysarthria; yet insight into his/her conditionmay remain. This pattern of progression relates to the cerebellar natureof this disease, with progression into the brain stem and eventually thecerebrum.

Fatal Familial Insomnia (FFI).

FFI typically presents in midlife (40s to 50s) with the insidious orsubacute onset of insomnia, initially manifest as a mild, then moresevere, reduction in overall sleep time. When sleep is achieved, vividdreams are common. A disturbance in autonomic function then emerges,which may manifest as elevated blood pressure, episodichyperventilation, excessive lacrimation, sexual and urinary tractdysfunction, and/or a change in basal body temperature. Signs ofbrainstem involvement, such as decreased ability to gaze upward, doublevision, jerky eye pursuit movements, or dysarthric speech may alsoappear in some individuals. With continued progression over the next fewmonths, individuals develop truncal and/or appendicular ataxia. Thespeed of thought processing may be reduced, as is common in subcorticaldementing states, and memory impairment may be variable; however,compared with other more prominent features of disease, cognitivecapacity is relatively spared until late in the course. Advancingdisease results in progressively greater loss of total sleep time,worsening ataxia, and more profound confusion, leading ultimately to anawake but stuporous state as death approaches. As with other forms ofprion disease, debilitation leading to feeding difficulties and loss ofairway protection is the most common immediate cause of death. Thetypical duration of disease is 12 to 16 months, with a range of a fewmonths to five years.

Other Prion Diseases.

About 10-15% of prion diseases are genetically transmissible, while theremainder occur from unknown risk factors or are acquired throughinfection with prions; these include sporadic Creutzfeldt-Jakob disease(sCJD), iatrogenic CJD (iCJD), variant CJD (vCJD), and sporadic fatalinsomnia (sFI). Kuru, a prion disease associated with the practice ofcannibalism in a primitive culture in New Guinea, is primarily ofhistorical significance.

sCJD.

The clinical and pathologic features of sCJD are the same as fCJD;however, the duration of disease is typically much shorter, on theaverage of six months or less, and the age at onset is later, typicallyafter age 60 years.

sFI.

The phenotype is the same as in FFI, including age at onset and durationof disease. sFI is much less common than FFI.

iCJD.

Diagnosis of this form of prion disease requires the identification orstrong association with administration of a biological extract or tissuecontaminated with prions. Such sources have included injections of humangrowth hormone contaminated with prions (used prior to 1980), improperlydecontaminated depth electrodes previously used in individuals with CJD,transplantation of corneas obtained from individuals with CJD, duramater grafts contaminated with prions, and various poorly documentedneurosurgical procedures.

vCJD.

This prion disease represents a relatively new strain of CJD acquired byingestion of beef or beef products contaminated with bovine spongiformencephalopathy (BSE), the prion disease of cattle (commonly known as madcow disease). The typical clinical picture is that of a young adult orteen who develops behavioral changes and/or pain in the lowerextremities that eventually lead to a progressive dementia with ataxiaand myoclonus. The course is about 1.5 years. The EEG is oftendiffusively slow rather than periodic, and the 14-3-3 CSF protein testis more often negative than positive. Neuropathology reveals spongiformchange spread diffusely throughout the brain and dense amyloid plaquedeposition surrounded by a halo of vacuolation described as “floridplaques.

SUMMARY

It has been determined by the present inventor that the gastrointestinaltract of dysautonomic individuals is impaired, and that the properlevels of pancreatic enzymes and/or their precursors including thezymogens and bicarbonate ions are not present in sufficient quantitiesto allow proper digestion. While that impairment is relevant to thedigestion of carbohydrates, fats and proteins, it is most specific andmost severe with respect to protein digestion. Accordingly, while notbeing bound by theory, the present inventor believes that many, if notall, dysautonomias have a GI component, and thus that dysautonomias mayactually have their etiology in gastrointestinal dysfunction. Forexample, with Guillaine-Barre syndrome, it is postulated that a GIpathogen is a causative factor in the formation of the Guillaine Barredysautonomia. Similarly, it has been found by the present inventor thatpopulations of autistic children suffer from GI disturbances and otherconditions which are dysautonomic in nature. In general, these findingsrepresent a possible link between the etiology of autism and autonomicdysfunction. Thus, the inventor believes that other dysautonomicconditions also have GI primary etiologies.

The symptoms of dysautonomic conditions, however, may have variousmanifestations due to the genetic makeup of the individuals sufferingfrom the conditions. Various gene sequences in the genetic code of theindividual will result in manifestation of certain diseases or symptomsthat are expressed uniquely in each individual. For example, if aminoacid pool deficits due to improper protein digestion andgastrointestinal dysfunction are manifested differently in differentindividuals, a “disease state” may appear different depending upon thegenetic makeup of the individual. Neurological expression may be allthat is seen in some individuals, whereas other manifestations maydemonstrate a hybrid of gastrointestinal dysfunction as well asneurological or other dysfunctions.

Accordingly, while not bound by theory, the present inventor believesthat prion diseases may have a dysautonomic component and that theetiology of prion diseases may be related to gastrointestinaldysfunction.

Given the above, it is a goal of the present disclosure to providetherapeutic methods and pharmaceutical compositions for the treatment ofthe symptoms of prion diseases. It is also a goal of the presentdisclosure to provide therapeutic methods and pharmaceuticalcompositions for the treatment of Pervasive Development Disorders suchas Autism, ADD, and ADHD, and for dysautonomias such as FamilialDysautonomia, Parkinson's, and Guillaine Barre Syndrome.

Another goal of the present disclosure is the provision ofpharmaceutical compositions for the treatment of the above disorders,wherein the compositions comprise one or more digestive enzymes, e.g.,one or more enzymes selected from amylases, proteases, cellulases,papaya, papain, bromelain, lipases, chymotrypsin, trypsin, andhydrolases. In some embodiments, the pharmaceutical compositions arelipid encapsulated.

Yet another goal of the present disclosure is to provide methods formaking the described pharmaceutical compositions using methods such as:direct compression, microencapsulation, lipid encapsulation, wetgranulation or other methods including the use of Prosolv® (silicifiedmicrocrystalline cellulose), and other known excipients and additives toaccomplish microencapsulation, lipid encapsulation, direct compression,wet or dry granulation or other suitable technology.

A further goal of the present disclosure is to provide means to deliverthe pharmaceutical compositions, which can include the use of rapiddissolution (rapid dissolve), time release, or other delivery methodsincluding oral, injection, patch, or other method. Further, the deliveryof the pharmaceutical compositions may be in the form of a tablet,capsule, sprinkles, sachet, or other oral delivery method.

An additional goal of the disclosure is to demonstrate the use of fecalchymotrypsin level as a biomarker for the presence of prion diseases, orthe likelihood of an individual to develop prion diseases.

Accordingly, provided herein is a method for treating one or moresymptoms associated with prion diseases in a patient diagnosed with aprion disease comprising administering to the patient a therapeuticallyeffective amount of a pharmaceutical composition comprising one or moredigestive enzymes. In some embodiments, the pharmaceutical compositionmay be lipid-encapsulated. In some embodiments, the one or moredigestive enzymes comprise one or more enzymes selected from the groupconsisting of proteases, amylases, celluloses, sucrases, maltases,papaya, papain, bromelain, hydrolases, and lipases. In some embodiments,the one or more digestive enzymes comprise one or more pancreaticenzymes. In some embodiments, the pharmaceutical composition comprisesone or more proteases, one or more lipases, and one or more amylases. Insome embodiments, the one or more proteases comprise chymotrypin andtrypsin.

The one or more digestive enzymes are, independently, derived from ananimal source, a microbial source, or a plant source, or aresynthetically prepared. In some embodiments, the animal source is a pig,e.g., a pig pancreas.

In some embodiments, the pharmaceutical composition comprises at leastone amylase, a mixture of proteases comprising chymotrypsin and trypsin,at least one lipase, and papain. In some embodiments, the pharmaceuticalcomposition further comprises papaya. In some embodiments, thepharmaceutical composition comprises, per dose: amylases from about10,000 to about 60,000 U.S.P; proteases from about 10,000 to about70,000 U.S.P; lipases from about 4,000 to about 30,000 U.S.P;chymotrypsin from about 2 to about 5 mg; trypsin from about 60 to about100 mg; papain from about 3,000 to about 10,000 USP units; and papayafrom about 30 to about 60 mg.

In some embodiments, the pharmaceutical composition comprises at leastone protease and at least one lipase, wherein the ratio of totalproteases to total lipases (in USP units) ranges from about 1:1 to about20:1. In some embodiments, the ratio of proteases to lipases ranges fromabout 4:1 to about 10:1.

In some embodiments, the one or more symptoms of prion diseases areselected from personality changes, psychiatric problems such asdepression, lack of coordination, and/or an unsteady gait. Patients alsomay experience involuntary jerking movements called myoclonus, unusualsensations, insomnia, confusion, or memory problems. In the later stagesof the disease, patients have severe mental impairment and lose theability to move or speak.

In some embodiments, the pharmaceutical composition is a dosageformulation selected from the group consisting of: pills, tablets,capsules, microcapsules, mini-capsules, time released capsules,mini-tabs, sprinkles, and a combination thereof.

Also provided is a method of diagnosing a patient comprising: obtaininga fecal sample from the patient; determining a level of chymotrypsinpresent in the fecal sample, wherein the determination is performed at30° C.; and diagnosing the patient as having a prion disease if thedetermined fecal chymotrypsin level is 8.4 U/gram or less and thepatient exhibits at least one symptom associated with a prion disease.In some embodiments, the fecal chymotrypsin level is between 8.4 and 4.2U/gram. In some embodiments, the fecal chymotrypsin level is less than4.2 U/gram. In some embodiments, the level of chymotrypsin present inthe fecal sample is determined using an enzymatic photospectrometrymethod. In some embodiments, the method further comprises administeringto the patient an effective amount of a pharmaceutical compositioncomprising one or more digestive enzymes if the patient is diagnosed ashaving a prion disease. In some embodiments, the method furthercomprises determining if the administration of the pharmaceuticalcomposition reduces or ameliorates one or more symptoms associated witha prion disease.

Also provided is a method of identifying a patient likely to benefitfrom administration of a pharmaceutical composition comprising one ormore digestive enzymes comprising: obtaining a fecal sample from thepatient; determining a level of chymotrypsin present in the fecalsample, wherein the determination is performed at 30° C.; andidentifying the patient as likely to benefit from administration of thepharmaceutical composition if the determined fecal chymotrypsin level is8.4 U/gram or less and the patient is diagnosed with a prion disease. Insome embodiments, the method further comprises determining if thepatient exhibits one or more symptoms of a prion disease. In someembodiments, the benefit comprises a reduction or amelioration of one ormore symptoms associated with a prion disease. In some embodiments, themethod further comprises administering to the patient an effectiveamount of a pharmaceutical composition comprising one or more digestiveenzymes.

Also provided is a pharmaceutical composition comprising one or moredigestive enzymes, wherein the one or more digestive enzymes comprise atleast one lipase and at least one protease, and wherein the ratio oftotal proteases to total lipases (in USP units) ranges from about 1:1 toabout 20:1. In some embodiments, the ratio of total proteases to totallipases ranges from about 4:1 to about 10:1. In some embodiments, thepharmaceutical composition is lipid encapsulated.

Also provided is a pharmaceutical composition comprising at least oneamylase, a mixture of proteases comprising chymotrypsin and trypsin, atleast one lipase, and papain. In some embodiments, the pharmaceuticalcomposition further comprises papaya. In some embodiments, the ratio oftotal proteases to total lipases ranges from about 1:1 to about 20:1.

The features and advantages described herein are not all-inclusive and,in particular, many additional features and advantages will be apparentto one of ordinary skill in the art in view of the drawings,specification, and claims. Moreover, it should be noted that thelanguage used in the specification has been principally selected forreadability and instructional purposes, and not to limit the scope ofthe inventive subject matter.

DETAILED DESCRIPTION

The present disclosure provides pharmaceutical compositions and methodsfor treating symptoms associated with CRPS, Pervasive DevelopmentDisorders, and Dysautonomias. The pharmaceutical compositions describedherein include one or more digestive enzymes, which are postulated bythe present inventor to assist in proper digest protein and thus toameliorate the gastrointestinal dysfunction that is associated with thedescribed disorders.

In certain embodiments, the pharmaceutical compositions may include oneor more digestive enzymes, wherein the one or more digestive enzymescomprise at least one lipase and at least one protease, and wherein theratio of total proteases to total lipases (in USP units) ranges fromabout 1:1 to about 20:1. In some cases, the ratio of total proteases tototal lipases ranges from about 4:1 to about 10:1.

In some cases, a pharmaceutical composition for use herein comprises atleast one amylase, at least one protease, and at least one lipase. Incertain embodiments, the pharmaceutical composition includes multipleproteases, including, without limitation, chymotrypsin and trypsin. Incertain embodiments, the composition can further include one or morehydrolases, papain, bromelain, papaya, celluloses, pancreatin, sucrases,and maltases.

The one or more enzymes can be independently derived from animal, plant,microbial, or synthetic sources. In some embodiments, the one or moreenzymes are derived from pig, e.g.: pig pancreas.

One exemplary formulation for the treatment of the symptoms of priondiseases is as follows:

Amylase 10,000-60,000 U.S.P

Protease 10,000-70,000 U.S.P

Lipase 4,000-30,000 U.S.P

Chymotrypsin 2-5 mg

Trypsin 60-100 mg

Papain 3,000-10,000 USP units/mg

Papaya 30-60 mg

Additional formulations comprising one or more digestive enzymes may beadvantageous including formulations in which the ratio of totalproteases to total lipases (in USP units) is from about 1:1 to about20:1. In some embodiments, the ratio of total proteases to total lipasesis from about 4:1 to about 10:1. Such formulations are useful fortreating symptoms of prion diseases as well as dysautonomias (e.g.,familial dysautonomia, Parkinson's, Guillaine-Barre Syndrome,Aromatic-L-amino acid decarboxylase deficiency, tetrahydrobiopterindeficiency, familial paranganglioma syndrome; multiple system atrophy,dysautonomic symptoms associated with tumors such as pheochromocytoma,chemodectoma, and neuroblastoma; neurally mediated syncope, and SIDS)and pervasive development disorders such as autism, ADHD, ADD, andAsperger's.

Patients below the age of 18 are typically given a dosage such that theformulation would deliver at least 5,000 USP units of protease and nomore than 10,000 USP units of lipase per kilogram weight of patient, perday. Beneficially, the formulation would deliver at least 5,000 USPunits of protease and no more than 7,500 USP units of lipase perkilogram weight of patient per day. Patients above the age of 18 aretypically given no less than 5,000 USP units of protease per kilogramweight of patient per day.

The dosage formulation may be administered by an oral preparationincluding, but not limited to, an encapsulated tablet, mini-tabs,microcapsule, mini-capsule, time released capsule, sprinkle or othermethodology. In one embodiment, the oral preparation is encapsulatedusing lipid. Alternatively, the oral preparation may be encapsulatedusing enteric coating or organic polymers. A formulation may also beprepared using Prosolv® technology, direct compression, dry granulation,wet granulation, and/or a combination of these methods.

Fecal chymotrypsin level is a sensitive, specific measure of proteolyticactivity, see e.g.: U.S. Pat. No. 6,660,831, incorporated by referenceherein. Normal levels of chymotrypsin are considered be greater than 8.4U/gram. Decreased values (less than 4.2 U/gram) suggest diminishedpancreatic output (pancreatic insufficiency), hypoacidity of the stomachor cystic fibrosis. Elevated chymotrypsin values suggest rapid transittime, or less likely, a large output of chymotrypsin from the pancreas.

For the fecal chymotrypsin test, a stool sample is collected from eachof the subjects. Each stool sample can be analyzed using an enzymaticphoto spectrometry analysis to determine the level of fecal chymotrypsinin the stool; in some cases the assay is performed at 30° C., see e.g.:U.S. Pat. No. 6,660,831, incorporated by reference herein.Alternatively, other methods, such as the colorimetric method, use ofsubstrates, use of assays, and/or any other suitable method may be usedto measure the fecal chymotrypsin levels. The levels of fecalchymotrypsin in the samples of the individuals having a prion diseaseare compared to the levels of fecal chymotrypsin in individuals notdiagnosed with a prion disease determine if the individuals having theprion disease would benefit from the administration of digestiveenzymes.

The foregoing description of the embodiments of the invention has beenpresented for the purposes of illustration and description. It is notintended to be exhaustive or to limit the invention to the precise formdisclosed. Many modifications and variations are possible in light ofthis disclosure. It is intended that the scope of the invention belimited not by this detailed description, but rather by the claimsappended hereto.

What is claimed is:
 1. A method for treating a prion disease in apatient diagnosed with the prion disease, the method comprisingadministering to the patient a pharmaceutical composition that comprisesdigestive enzymes, wherein the digestive enzymes comprise a protease, anamylase and a lipase, whereby one or more symptoms of the prion diseaseare treated; wherein said patient diagnosed with the prion disease has asubnormal level of fecal chymotrypsin, wherein the one or more symptomscomprise a personality change, a psychiatric problem, lack ofcoordination, unsteady gait, myoclonus, an unusual sensation, insomnia,confusion, a memory problem, severe mental impairment, loss of theability to move or speak, or a combination thereof.
 2. The method ofclaim 1, wherein the pharmaceutical composition further comprises one ormore enzymes selected from the group consisting of a cellulase, asucrase, and a maltase.
 3. The method of claim 1, wherein the digestiveenzymes are pancreatic enzymes.
 4. The method of claim 1, wherein theprotease is chymotrypsin or trypsin.
 5. The method of claim 1, whereinthe digestive enzymes are animal enzymes, microbial source enzymes,plant enzymes, or are synthetically prepared enzymes.
 6. The method ofclaim 1, wherein the amylase is present in the pharmaceuticalcomposition in an amount of from about 10,000 to about 60,000 UnitedStates Pharmacopeia (U.S.P) units per dose.
 7. The method of claim 1,wherein the protease and the lipase in the pharmaceutical composition inU.S.P. units are present in a ratio of from about 1:1 to about 20:1. 8.The method of claim 7, wherein the ratio of protease to lipase is fromabout 4:1 to about 10:1.
 9. The method of claim 1, wherein thepharmaceutical composition is a dosage formulation selected from thegroup consisting of a pill, a tablet, a capsule, a microcapsule, amini-capsule, a time released capsule, a mini-tab, a sprinkle, and acombination thereof.
 10. The method of claim 1, wherein the subnormallevel of fecal chymotrypsin level is 8.4 U/gram or less.
 11. The methodof claim 1, wherein the protease is present in the pharmaceuticalcomposition in an amount of from about 10,000 to about 70,000 U.S.Punits per dose.
 12. The method of claim 1, wherein the lipase is presentin the pharmaceutical composition in an amount of from about 4,000 toabout 30,000 U.S.P. units per dose.
 13. The method of claim 4, whereinthe protease is chymotrypsin, and wherein the chymotrypsin is present inthe pharmaceutical composition in an amount of from about 2 to about 5mg per dose.
 14. The method of claim 4, wherein the protease is trypsin,and wherein the trypsin is present in the pharmaceutical composition inan amount of from about 60 to about 100 mg per dose.